A method and apparatus of transmitting a reference signal in a wireless communication system is provided. Demodulation Reference Signals (DMRSs) for a plurality of respective antennas is generated. The DMRSs are mapped to a resource region, and transmitted through the respective corresponding antennas. The DMRSs are multiplexed using at least one of frequency division multiplexing (FDM), code division multiplexing (CDM), and time division multiplexing (TDM) methods and mapped in the resource region. Also, a position of an orthogonal frequency division multiplexing (OFDM) symbol to which the DMRSs are mapped in the resource region is an OFDM symbol to which a physical downlink control channel (PDCCH) and a legacy cell-specific reference signal (CRS) are not mapped.